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Justin Kennedy Dissertation Defense for the Degree of Doctorate in Philosophy

Efficient Algorithms for SNP Genotype Data Analysis using Hidden Markov Models of Haplotype Diversity. Justin Kennedy Dissertation Defense for the Degree of Doctorate in Philosophy Computer Science & Engineering Department University of Connecticut. Outline. Introduction

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Justin Kennedy Dissertation Defense for the Degree of Doctorate in Philosophy

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  1. Efficient Algorithms for SNP Genotype Data Analysis using HiddenMarkov Models of Haplotype Diversity Justin Kennedy Dissertation Defense for the Degree of Doctorate in Philosophy Computer Science & Engineering Department University of Connecticut

  2. Outline • Introduction • Hidden Markov Models of Haplotype Diversity • Genotype Error Detection using Hidden Markov Models of Haplotype Diversity • Imputation-based Local Ancestry Inference in Admixed Populations • Single Individual Genotyping from Low-Coverage Sequencing Data • Conclusion

  3. Introduction-Single Nucleotide Polymorphisms • Main form of variation between individual genomes: Single Nucleotide Polymorphisms (SNPs) • High density in the human genome:  1.3x107out of 3109 base pairs • Vast majority bi-allelic  0/1 encoding (major/minor resp.) • Haplotype: description of SNP alleles on a chromosome • 0/1 vector: 0 for major allele, 1 for minor … ataggtccCtatttcgcgcCgtatacacgggActata … … ataggtccGtatttcgcgcCgtatacacgggTctata … … ataggtccCtatttcgcgcGgtatacacgggTctata …

  4. 011000110 001100010 012100120 two haplotypes per individual + genotype Genotype Error Detection-SNP Genotypes • Diploid: two haplotypes for each chromosome • One inherited from mother and one from father • Multilocus Genotype: description of alleles on both chromosomes • 0/1/2 vector: 0 (2) - both chromosomes contain the major (minor) allele; 1 - the chromosomes contain different alleles • SNP Genotypes are critical to Disease-Gene Mapping

  5. Introduction- Why SNP Genotypes? • SNPs are the genetic marker of choice for genome wide association studies (GWASs) • GWAS: Method for discovering disease associated genes by typing a dense set of markers in large numbers of cases and controls followed by a statistical test of association. • Ongoing GWASs generate a deluge of genotype data • Genetic Association Information Network (GAIN): 6 studies totaling 19,000 individuals typed at 500,000 to 940,000 SNP loci • Wellcome Trust Case-Control Consortium (WTCCC): 7 studies totaling 17,000 individuals typed at 500,000 SNPs • WTCCC2: hundreds of thousands of individuals covering over a million SNPs!

  6. Introduction-Computational Challenges to Disease Gene Mapping • Genotype error detection: Genotyping errors can decrease statistical power and invalidate statistical tests for disease association based on haplotypes. • Local ancestry Inference: Accurate estimates of local ancestry surrounding disease-associated loci are a critical step in admixture mapping. • Accurate SNP Genotyping from new sequencing technologies: Accurate determination of both alleles at variable loci is essential, and is limited by coverage depth due to random nature of shotgun sequencing.

  7. Outline • Introduction • Hidden Markov Models of Haplotype Diversity • Genotype Error Detection using Hidden Markov Models of Haplotype Diversity • Imputation-based Local Ancestry Inference in Admixed Populations • Single Individual Genotyping from Low-Coverage Sequencing Data • Conclusion

  8. Haplotype structure in panmictic populations

  9. HMM of haplotype diversity n = 5 (# SNPs) • Similar models proposed in [Schwartz 04, Rastas et al. 08, Kennedy et al. 07, Kimmel&Shamir 05, Scheet&Stephens 06,…] • Captures Linkage Disequilibrium (LD) k= 4 (# founders)

  10. Graphical model representation … F1 F2 Fn • Random variables for each locus i (i=1..n) • Fi = founder haplotype at locus i; values between 1 and k • Hi = observed allele at locus i; values: 0 (major) or 1 (minor) • Model training, based on Baum-Welch algorithm, using: • Reference haplotypes from population panel (e.g. Hapmap), or • Haplotypes from phased genotype using ENT software • Given haplotype h, P(H=h|M) can be computed in O(nk2) using a forward algorithm. H1 H2 Hn

  11. Factorial HMM for genotype data … F1 F2 Fn H1 H2 Hn … F'1 F'2 F'n H'1 H'2 H'n G1 G2 Gn • Random variable for each locus i (i=1..n) • Gi= genotype at locus i; values: 0/1/2 (major hom./het./minor hom.) • Given multilocus genotype g, P(g|M) can be computed in O(nk4) using a forward algorithm.

  12. HMM Based Genotype Imputation • Probability of observing genotype at locus i given the known multilocus genotype with missing data at i: •  Giis imputed as:

  13. Forward-backward computation Fi … … Hi F’i … … H’i Gi

  14. Forward-backward computation Fi … … Hi F’i … … H’i Gi

  15. Forward-backward computation Fi … … Hi F’i … … H’i Gi

  16. Forward-backward computation Fi … … Hi F’i … … H’i Gi

  17. Runtime reduced to O(nk3) by reusing common terms: where Runtime • Direct recurrences for computing forward probabilities O(nk4):

  18. Speed-up: PopTreeTrie

  19. Outline • Introduction • Hidden Markov Models of Haplotype Diversity • Genotype Error Detection using Hidden Markov Models of Haplotype Diversity • Motivation • Likelihood Sensitivity Approach to Error Detection • Hidden Markov Model of Haplotype Diversity • Efficiently Computable Likelihood functions • Experimental Results • Imputation-based Local Ancestry Inference in Admixed Populations • Single Individual Genotyping from Low-Coverage Sequencing Data • Conclusion

  20. Genotype Errors- Motivation • A real problem despite advances in genotyping technology • [Zaitlen et al. 2005] found 1.1% inconsistencies among the 20 million dbSNP genotypes typed multiple times • 1% errors decrease power by 10-50% for linkage, and by 5-20% for association • Error types • Easily Detectable errors • Systematic errors (e.g., assay failure) detected by HWE test [Hosking et al. 2004] • For pedigree data some errors detected as Mendelian Inconsistencies (MIs) • E.g. Only ~30% detectable as MIs for trios [Gordon et al. 1999] • Undetected errors • Methods for handling undetected errors: • Improved genotype calling algorithms • Improved modeling in analysis methods • Separate error detection step • Detected errors can be retyped, imputed, or ignored in downstream analysis

  21. Mother Father 0 1 2 1 0 2 0 2 2 1 0 2 0 0 0 1 0 1 H3 0 1 1 1 0 0 H4 0 1 1 1 0 0 H1 0 1 0 1 0 1 H2 Child 0 2 2 1 0 2 0 1 1 1 0 0 H1 0 0 0 1 0 1 H3 Likelihood of best phasing for original trio T Genotype Error Detection- Likelihood Sensitivity Approach to Error Detection [Becker et al. 06]

  22. Mother Father 0 1 2 1 0 2 0 2 2 1 0 2 0 0 0 1 0 0 H’ 3 0 1 1 1 0 1 H’ 4 0 1 0 1 0 1 H’1 0 1 1 1 0 0 H’2 Child 0 2 2 1 0 2 0 1 0 1 0 1 H’ 1 0 0 0 1 0 0 H’ 3 Likelihood of best phasing for modified trio T’ Genotype Error Detection- Likelihood Sensitivity Approach to Error Detection [Becker et al. 06] ? Likelihood of best phasing for original trio T

  23. Mother Father 0 1 2 1 0 2 0 2 2 1 0 2 Child 0 2 2 1 0 2 Genotype Error Detection- Likelihood Sensitivity Approach to Error Detection [Becker et al. 06] Mother Father 0 1 2 1 0 2 0 2 2 1 0 2 Child 0 2 2 1 0 2 ? • Large change in likelihood suggests likely error • Flag genotype as an error if L(T’)/L(T) > R, where R is the detection threshold (e.g., R=104)

  24. Mother …201012 1 02210... Father …201202 2 10211... Child …000120 2 21021... Genotype Error Detection- Likelihood Sensitivity Approach to Error Detection • [Becker et al. 06] Implementation in FAMHAP Software • Window-based algorithm • For each window including the SNP under test, generate list of H most frequent haplotypes (default H=50) • Find most likely trio phasings by pruned search over the H4 quadruples of frequent haplotypes • Flag genotype as an error if L(T’)/L(T) > R for at least one window

  25. Genotype Error Detection- Limitations of FAMHAP • Unbounded list of haplotypes (H=4n) is hard to compute • Truncating H may lead to sub-optimal phasings and inaccurate L(T) values • False positives caused by nearby errors (due to the use of multiple short windows) Our approach: • HMM of haplotype frequencies  all haplotypes represented + no need for short windows • Alternate likelihood functions  scalable runtime

  26. Trio-Based HMM of haplotype diversity F1 F2 … Fn F1 F2 … Fn H1 H2 Hn H1 H2 Hn … F'n F‘1 F‘2 F’1 F’2 … F’n H'1 H'2 H'n H’1 H’2 H’n GM1 GM2 GMn GF1 GF2 GFn GC1 GC2 GCn

  27. Genotype Error Detection- Alternate Likelihood Functions • Viterbi probability (ViterbiProb): Maximum prob. of a set of 4 HMM paths that emit 4 haplotypes compatible with the trio. • Probability of Viterbi Haplotypes (ViterbiHaps): Obtain the path of the 4 Viterbi haplotypes, then then take product of these individual haplotype probabilities using forward (again). • Total Trio Probability (TotalProb): Total probability P(T) that the HMM emits four haplotypes that explain trio T along all possible 4-tuples of paths.

  28. Genotype Error Detection- Speed Ups from reuse of common terms • Straight-forward approach run time: • For a fixed trio, ViterbiProb/TotalProb paths can be found using a 4-path version of Viterbi’s/Forward algorithm in time • For ViterbiHaps, additional traceback to compute probabilities: • K3 speed-up by reuse of common terms: per trio • Likelihoods of all 3n modified trios computed using forward-backward algorithm, • ViterbiProb/TotalProb for m trios: • ViterbiHaps:

  29. Genotype Error Detection- Comparison of Likelihood Functions • 35 SNPs • 551 trios • [Becker 06] • 1% err. rate Sensitivity=TP/(TP+TN) False Positive rate = 1 - TN/(FP+TN)

  30. Genotype Error Detection-“Combined” Detection Method • Compute 4 likelihood ratios • Trio • Mother-child duo • Father-child duo • Child (unrelated) • Flag as error if all ratios are above detection threshold

  31. 35 SNPs • 551 trios • [Becker 06] • 1% err. rate Genotype Error Detection- Comparison with FAMHAP

  32. Outline • Introduction • Hidden Markov Models of Haplotype Diversity • Genotype Error Detection using Hidden Markov Models of Haplotype Diversity • Imputation-based Local Ancestry Inference in Admixed Populations • Motivation • Factorial HMM of genotype data • Algorithms for genotype imputation and ancestry inference • Experimental results • Single Individual Genotyping from Low-Coverage Sequencing Data • Conclusion

  33. Introduction- Population admixture http://www.garlandscience.co.uk/textbooks/0815341857.asp?type=resources

  34. Introduction- Motivation: Admixture mapping Patterson et al, AJHG 74:979-1000, 2004

  35. Introduction- Local ancestry inference problem • Given: • Reference haplotypes for all ancestral populations to be studied • Whole-genome SNP genotype data for extant individual • Find: • Allele ancestries at each SNP locus Reference haplotypes 1110001?0100110010011001111101110111?1111110111000 11100011010011001001100?100101?10111110111?0111000 11110010011001101001110010110101011111011110111000 1110001001000100111110001111011100111?111110111000 011101100110011011111100101101110111111111?0110000 11100010010001001111100010110111001111111110110000 011?001?011001101111110010?10111011111111110110000 11100110010001001111100011110111001111111110111000 1110001?0100110010011001111101110111?1111110111000 11100011010011001001100?100101?10111110111?0111000 11110010011001101001110010110101011111011110111000 1110001001000100111110001111011100111?111110111000 011101100110011011111100101101110111111111?0110000 11100010010001001111100010110111001111111110110000 011?001?011001101111110010?10111011111111110110000 11100110010001001111100011110111001111111110111000 1110001?0100110010011001111101110111?1111110111000 11100011010011001001100?100101?10111110111?0111000 11110010011001101001110010110101011111011110111000 1110001001000100111110001111011100111?111110111000 011101100110011011111100101101110111111111?0110000 11100010010001001111100010110111001111111110110000 011?001?011001101111110010?10111011111111110110000 11100110010001001111100011110111001111111110111000 Inferred local ancestry rs11095710 P1 P1 rs11117179 P1 P1 rs11800791 P1 P1 rs11578310 P1 P2 rs1187611 P1 P2 rs11804808 P1 P2 rs17471518 P1 P2 ... SNP genotypes rs11095710 T T rs11117179 C T rs11800791 G G rs11578310 G G rs1187611 G G rs11804808 C C rs17471518 A G ...

  36. Introduction- Previous work Two main classes of methods for SNP Ancestry Inference HMM-based (exploit LD): SABER [Tang et al 06], SWITCH [Sankararaman et al 08a], HAPAA [Sundquist et al. 08], … Window-based (unlinked SNP Data): LAMP [Sankararaman et al 08b], WINPOP [Pasaniuc et al. 09] Limitations Poor accuracy when ancestral populations are closely related (e.g. Japanese and Chinese) Methods based on unlinked SNPs outperform methods that model LD!

  37. Factorial HMM for genotype data in a window with known local ancestry … F1 F2 Fn H1 H2 Hn … F'1 F'2 F'n H'1 H'2 H'n G1 G2 Gn

  38. Fixed-window version: pick ancestry that maximizes the average posterior probability of the SNP genotypes within a fixed-size window centered at the locus Observations: The local ancestry of a SNP locus is typically shared with neighboring loci. Small Window sizes may not provide enough information Large Window sizes may violate local ancestry property for neighboring loci Imputation-based ancestry inference

  39. Window size effect N=2,000 g=7 =0.2 n=38,864 r=10-8

  40. Multi-window version: Weighted voting over window sizes between 200-3000, with window weights proportional to average posterior probabilities Imputation-based ancestry inference

  41. Comparison with other methods % of correctly recovered SNP ancestries N=2,000 g=7 =0.2 n=38,864 r=10-8

  42. Untyped SNP imputation error rate in admixed individuals N=2,000 g=7 =0.5 n=38,864 r=10-8

  43. Genotype Imputation- Accuracy with number founders/runtime • 5835 SNPs • 2502 unrelated (CEU) • [IMAGE] • 9% imputed (535 SNPs)

  44. Number of founders effect on Ancestry inference CEU-JPT N=2,000 g=7 =0.2 n=38,864 r=10-8

  45. Outline • Introduction • Hidden Markov Models of Haplotype Diversity • Genotype Error Detection using Hidden Markov Models of Haplotype Diversity • Imputation-based Local Ancestry Inference in Admixed Populations • Single Individual Genotyping from Low-Coverage Sequencing Data • Motivation • Single SNP Calling Algorithms • HF-HMM Overview • Multilocus HMM Calling Algorithm • Experimental Results • Conclusion

  46. Low Coverage Genotyping-Next Generation Sequencing (NGS) • By several orders of magnitude, NGS delivers higher throughput of sequencing reads compared to older technologies (e.g. Sanger sequencing) Roche/454 FLX Titanium ~1M reads 400bp avg. 400-600Mb / run (10h) Illumina Genome Analyzer IIx ~100-300M reads/pairs 35-100bp 4.5-33 Gb / run (2-10 days) ABI SOLiD 3 plus ~500M reads/pairs 35-50bp 25-60Gb / run (3.5-14 days)

  47. Low Coverage Genotyping-NGS Applications and Challenges NGS is enabling many applications, including personal genomics ~$100 million for the Sanger-sequenced Venter genome [Levy et al 07] ~$1 million for sequencing James Watson genome [Wheeler et al 08] using 454 technology. ~$50K human sequencing now available Thousands more individual genomes to be sequenced as part of 1000 Genomes Project Challenges: Sequencing requires accurate determination of genetic variation (e.g. SNPs) Accuracy is limited by coverage depth due to random nature of shotgun sequencing For the Venter and Watson genomes (both sequenced at ~7.5x average coverage), only 75-80% accuracy achieved for sequencing based calls of heterozygous SNPs [Levy et al 07, Wheeler et al 08]. [Wheeler et al 08] use hypothesis testing based on binomial distribution

  48. Low Coverage Genotyping-Do Heuristic Inputs Help? [Wendl&Wilson 08] predict that 21x coverage is required for sequencing of samples based on the assumption that “neglects any heuristic inputs” We propose methods incorporating two additional sources of information: Quality scores reflecting uncertainty in sequencing data Linkage disequilibrium (LD) information and allele frequencies extracted from reference panels such as Hapmap

  49. Low Coverage Genotyping-Pipeline for Single Genotype Calling

  50. Single SNP Genotyping- Basic Notations • Biallelic SNPs: 0 = major allele, 1 = minor allele (reads with non-reference alleles are discarded) • SNP genotypes: 0/2 = homozygous major/minor, 1=heterozygous • Read set ri describes the mapped reads for each SNP i Mapped reads with allele 0 012100120 Inferred genotypes Mapped reads with allele 1 Sequencing errors

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